Abstract:
:The gltA gene, encoding Sinorhizobium meliloti 104A14 citrate synthase, was isolated by complementing an Escherichia coli gltA mutant. The S. meliloti gltA gene was mutated by inserting a kanamycin resistance gene and then using homologous recombination to replace the wild-type gltA with the gltA::kan allele. The resulting strain, CSDX1, was a glutamate auxotroph, and enzyme assays confirmed the absence of a requirement for glutamate. CSDX1 did not grow on succinate, malate, aspartate, pyruvate, or glucose. CSDX1 produced an unusual blue fluorescence on medium containing Calcofluor, which is different from the green fluorescence found with 104A14. High concentrations of arabinose (0.4%) or succinate (0. 2%) restored the green fluorescence to CSDX1. High-performance liquid chromatography analyses showed that CSDX1 produced partially succinylated succinoglycan. CSDX1 was able to form nodules on alfalfa, but these nodules were not able to fix nitrogen. The symbiotic defect of a citrate synthase mutant could thus be due to disruption of the infection process or to the lack of energy generated by the tricarboxylic acid cycle.
journal_name
J Bacterioljournal_title
Journal of bacteriologyauthors
Mortimer MW,McDermott TR,York GM,Walker GC,Kahn MLdoi
10.1128/JB.181.24.7608-7613.1999keywords:
subject
Has Abstractpub_date
1999-12-01 00:00:00pages
7608-13issue
24eissn
0021-9193issn
1098-5530journal_volume
181pub_type
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更新日期:1979-04-01 00:00:00
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更新日期:2007-07-01 00:00:00